Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 3.870
Filtrar
1.
Int J Mol Sci ; 22(4)2021 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-33669854

RESUMO

Changes that occur within oviducts after fertilization are dependent on post-ovulation events, including oocyte-oviduct interactions. Although general processes are well-defined, the molecular basis are poorly understood. Recently, new marker genes involved in 'cell development', 'cell growth', 'cell differentiation' and 'cell maturation' processes have been identified in porcine oocytes. The aim of the study was to assess the expression profile of genes in primary in vitro cultured oviductal epithelial cells (OECs), clustered in Gene Ontology groups which enveloped markers also identified in porcine oocytes. OECs (from 45 gilts) were surgically removed and cultured in vitro for ≤ 30 days, and then subjected to molecular analyses. The transcriptomic and proteomic profiles of cells cultured during 7, 15 and 30 days were investigated. Additionally, morphological/histochemical analyzes were performed. The results of genes expression profiles were validated after using RT-qPCR. The results showed a significant upregulation of UNC45B, NOX4, VLDLR, ITGB3, FMOD, SGCE, COL1A2, LOX, LIPG, THY1 and downregulation of SERPINB2, CD274, TXNIP, CELA1, DDX60, CRABP2, SLC5A1, IDO1, ANPEP, FST. Detailed knowledge of the molecular pathways occurring in the OECs and the gametes that contact them may contribute both to developments of basic science of physiology, and new possibilities in advanced biotechnology of assisted reproduction.


Assuntos
Células Epiteliais/metabolismo , Regulação da Expressão Gênica , Oócitos/metabolismo , Oviductos/citologia , Animais , Diferenciação Celular/genética , Forma Celular/genética , Células Cultivadas , Regulação para Baixo/genética , Feminino , Ontologia Genética , Redes Reguladoras de Genes , Marcadores Genéticos , Transdução de Sinais/genética , Suínos , Transcriptoma , Regulação para Cima/genética
2.
Nat Commun ; 12(1): 1251, 2021 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-33623007

RESUMO

Dysfunction of embryo transport causes ectopic pregnancy which affects approximately 2% of conceptions in the US and Europe, and is the most common cause of pregnancy-related death in the first trimester. Embryo transit involves a valve-like tubal-locking phenomenon that temporarily arrests oocytes at the ampullary-isthmic junction (AIJ) where fertilisation occurs, but the mechanisms involved are unknown. Here we show that female mice lacking the orphan adhesion G-protein coupled receptor Adgrd1 are sterile because they do not relieve the AIJ restraining mechanism, inappropriately retaining embryos within the oviduct. Adgrd1 is expressed on the oviductal epithelium and the post-ovulatory attenuation of tubal fluid flow is dysregulated in Adgrd1-deficient mice. Using a large-scale extracellular protein interaction screen, we identified Plxdc2 as an activating ligand for Adgrd1 displayed on cumulus cells. Our findings demonstrate that regulating oviductal fluid flow by Adgrd1 controls embryo transit and we present a model where embryo arrest at the AIJ is due to the balance of abovarial ciliary action and the force of adovarial tubal fluid flow, and in wild-type oviducts, fluid flow is gradually attenuated through Adgrd1 activation to enable embryo release. Our findings provide important insights into the molecular mechanisms involved in embryo transport in mice.


Assuntos
Líquidos Corporais/fisiologia , Embrião de Mamíferos/metabolismo , Oviductos/metabolismo , Receptores Acoplados a Proteínas-G/metabolismo , Reologia , Animais , Membrana Celular/metabolismo , Membrana Celular/ultraestrutura , Cílios/metabolismo , Cílios/ultraestrutura , Células do Cúmulo/metabolismo , Epitélio/metabolismo , Feminino , Genótipo , Infertilidade Feminina/metabolismo , Infertilidade Feminina/patologia , Ligantes , Masculino , Camundongos , Modelos Biológicos , Músculos/metabolismo , Mutação/genética , Oviductos/patologia , Oviductos/ultraestrutura , Regiões Promotoras Genéticas/genética , Ligação Proteica , Receptores de Superfície Celular/metabolismo , Receptores Acoplados a Proteínas-G/deficiência
3.
Zoolog Sci ; 38(1): 8-19, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33639713

RESUMO

Foam nests of frogs are natural biosurfactants that contain potential compounds for biocompatible materials, Drug Delivery System (DDS), emulsifiers, and bioremediation. To elucidate the protein components in the foam nests of Rhacophorus arboreus, which is an endemic Japanese frog species commonly seen during the rainy season, we performed amino acid analysis, SDS-PAGE electrophoresis, and matrix-assisted laser desorption/ionization mass spectrometry using intact foam nests. Many proteins were detected in these foam nests, ranging from a few to several hundred kDa, with both essential and non-essential amino acids. Next, we performed transcriptome analysis using a next-generation sequencer on total RNAs extracted from oviducts before egg-laying. The soluble foam nests were purified by LC-MS and analyzed using Edman degradation, and the identified N-terminal sequences were matched to the transcriptome data. Four proteins that shared significant sequence homologies with extracellular superoxide dismutase of Nanorana parkeri, vitelline membrane outer layer protein 1 homolog of Xenopus tropicalis, ranasmurfin of Polypedates leucomystax, and alpha-1-antichymotrypsin of Sorex araneus were identified. Prior to purification of the foam nests, they were treated with both a reducing reagent and an alkylating agent, and LC-MS/ MS analyses were performed. We identified 22 proteins in the foam nests that were homologous with proteinase inhibitors, ribonuclease, glycoproteins, antimicrobial protein and barrier, immunoglobulin-binding proteins, glycoprotein binding protein, colored protein, and keratin-associated protein. The presence of these proteins in foam nests, along with small molecules, such as carbohydrates and sugars, would protect them against microbial and parasitic attack, oxidative stress, and a shortage of moisture.


Assuntos
Anuros/metabolismo , Comportamento de Nidação/fisiologia , Oviductos/metabolismo , Proteoma , Animais , Anuros/genética , Feminino , Perfilação da Expressão Gênica
4.
Zoolog Sci ; 38(1): 20-25, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33639714

RESUMO

Oviparous, ovoviviparous and viviparous reproduction are interesting subjects for understanding animals' evolutionary pathways and adaptation to their life history and habitat conditions. In this study, we examined the reproductive mode of the ovoviviparous mayfly Cloeon dipterum, particularly comparing embryogenesis between hand-pairing and unmated females' common oviduct. Our study suggested that the high developmental rate of C. dipterum observed in a recent study could be ascribed to their absorption of unfertilized eggs. The developmental rates of hand-paired females were almost 100%, while their egg-bearing numbers were lower than those of virgin females. Thus, such reduced egg numbers suggest the maternal absorption of unfertilized eggs. This trait is thought to have evolved with the ovoviviparous characteristics of C. dipterum. We identified the basis of the irregularity of this species exhibiting such a high (i.e., 100%) developmental rate in our previous recent study.


Assuntos
Desenvolvimento Embrionário , Ephemeroptera/embriologia , Ephemeroptera/fisiologia , Óvulo , Animais , Tamanho Corporal , Embrião não Mamífero , Feminino , Masculino , Oviductos/fisiologia , Ovoviviparidade
5.
Int J Mol Sci ; 22(2)2021 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-33430094

RESUMO

Extracellular vesicles (EVs) have been isolated from follicular (FF) and ampullary oviduct fluid (AOF), using different isolation methods. However, it is not clear whether different purification methods can affect the functionality of resulting EVs. Here, we compared two methods (OptiPrep™ density gradient ultracentrifugation (ODG UC) and single-step size exclusion chromatography (SEC) (qEV IZON™ single column)) for the isolation of EVs from bovine FF and AOF. Additionally, we evaluated whether the addition of EVs derived either by ODG UC or SEC from FF or AOF during oocyte maturation would yield extra benefits for embryo developmental competence. The characterization of EVs isolated using ODG UC or SEC from FF and AOF did not show any differences in terms of EV sizes (40-400 nm) and concentrations (2.4 ± 0.2 × 1012-1.8 ± 0.2 × 1013 particles/mL). Blastocyst yield and quality was higher in groups supplemented with EVs isolated from FF and AOF by ODG UC, with higher total cell numbers and a lower apoptotic cell ratio compared with the other groups (p < 0.05). Supplementing in vitro maturation media with EVs derived by ODG UC from AOF was beneficial for bovine embryo development and quality.


Assuntos
Técnicas de Cultura Embrionária , Desenvolvimento Embrionário/genética , Vesículas Extracelulares/metabolismo , Oogênese/efeitos dos fármacos , Animais , Blastocisto/efeitos dos fármacos , Bovinos , Centrifugação com Gradiente de Concentração , Meios de Cultivo Condicionados , Feminino , Líquido Folicular/química , Líquido Folicular/metabolismo , Células Ciliadas da Ampola/química , Células Ciliadas da Ampola/metabolismo , Humanos , Oviductos/efeitos dos fármacos
6.
Cell Prolif ; 54(3): e12996, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33496365

RESUMO

OBJECTIVES: The female reproductive tract comprises several different cell types. Using three representative Cre systems, we comparatively analysed the phenotypes of Dgcr8 conditional knockout (cKO) mice to understand the function of Dgcr8, involved in canonical microRNA biogenesis, in the female reproductive tract. MATERIALS AND METHODS: Dgcr8f/f mice were crossed with Ltficre/+ , Amhr2cre/+ or PRcre/+ mice to produce mice deficient in Dgcr8 in epithelial (Dgcr8ed/ed ), mesenchymal (Dgcr8md/md ) and all the compartments (Dgcr8td/td ) in the female reproductive tract. Reproductive phenotypes were evaluated in Dgcr8 cKO mice. Uteri and/or oviducts were used for small RNA-seq, mRNA-seq, real-time RT-PCR, and/or morphologic and histological analyses. RESULT: Dgcr8ed/ed mice did not exhibit any distinct defects, whereas Dgcr8md/md mice showed sub-fertility and oviductal smooth muscle deformities. Dgcr8td/td mice were infertile due to anovulation and acute inflammation in the female reproductive tract and suffered from an atrophic uterus with myometrial defects. The microRNAs and mRNAs related to immune modulation and/or smooth muscle growth were systemically altered in the Dgcr8td/td uterus. Expression profiles of dysregulated microRNAs and mRNAs in the Dgcr8td/td uterus were different from those in other genotypes in a Cre-dependent manner. CONCLUSIONS: Dgcr8 deficiency with different Cre systems induces overlapping but distinct phenotypes as well as the profiles of microRNAs and their target mRNAs in the female reproductive tract, suggesting the importance of selecting the appropriate Cre driver to investigate the genes of interest.


Assuntos
Proteínas de Ligação a RNA/genética , Reprodução/genética , Útero/patologia , Animais , Feminino , Integrases/metabolismo , Integrases/farmacologia , Camundongos Knockout , MicroRNAs/genética , Oviductos/crescimento & desenvolvimento , Oviductos/metabolismo , Proteínas de Ligação a RNA/metabolismo , Reprodução/fisiologia , Útero/metabolismo
7.
Anim Sci J ; 91(1): e13456, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32926548

RESUMO

In this study, we investigated whether bovine oviducts and endometria produce anti-Müllerian hormone (AMH) (for paracrine and autocrine signaling). Reverse transcription-polymerase chain reaction and western blotting detected AMH expression in oviductal and endometrial specimens. Immunohistochemistry revealed robust AMH expression in the ampulla and isthmus epithelia, and the glandular and luminal endometrial epithelia (caruncular endometria). AMH mRNA (measured by real-time PCR) and protein expression in these layers did not significantly differ among estrous phases in adult Japanese Black (JB) heifers (p > .1). Furthermore, the expression in these layers also did not differ among Holstein cows (93.8 ± 5.8 months old), JB heifers (25.5 ± 0.4 months old), and JB cows (97.9 ± 7.9 months old). We also compared AMH concentrations in the oviduct and uterine horn fluids among the three groups (measured by immunoassays). Interestingly, the AMH concentration in the oviduct fluid, but not in the uterine horn fluid, of Holstein cows was lower than those in JB heifers and cows (p < .05). Therefore, bovine oviducts and endometria express AMH and likely secrete it into the oviduct and uterine fluids.


Assuntos
Hormônio Antimülleriano/genética , Hormônio Antimülleriano/metabolismo , Endométrio/metabolismo , Células Epiteliais/metabolismo , Expressão Gênica , Oviductos/metabolismo , Animais , Bovinos , Endométrio/citologia , Ciclo Estral/metabolismo , Feminino , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Útero/metabolismo
8.
Mol Immunol ; 127: 21-30, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32905905

RESUMO

Estrogen has known anti-inflammatory effects, but the mechanism whereby 17ß-estradiol (E2) protects oviduct sheep epithelial cells from inflammation remains unknown. In this study, we detected the E2 synthetase and E2 nuclear and membrane receptors in sheep oviducts, primarily in epithelial cells. Using lipopolysaccharide (LPS)-stimulated sheep oviduct epithelial cells as an in vitro inflammation model, we demonstrated that E2 attenuates the expression of inflammatory factors in a concentration-response manner. E2 also inhibited the LPS-stimulated phosphorylation of p38 MAPK and NF-κB p65 but did not reduce the phosphorylation of JNK and ERK 1/2. This attenuation was partially antagonized by an intracellular estrogen antagonist that was involved in genomic regulation and enhanced by a G protein-coupled estrogen receptor agonist that was involved in nongenomic cellular modulation. These results suggest that E2 has an inhibitory effect on LPS-induced oviduct epithelial cell inflammation in sheep, which is mediated by the downstream regulatory effects of estrogen receptors.


Assuntos
Células Epiteliais/patologia , Estradiol/farmacologia , Inflamação/patologia , Oviductos/patologia , Substâncias Protetoras/farmacologia , Animais , Aromatase/metabolismo , Citocinas/genética , Citocinas/metabolismo , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Feminino , Fulvestranto/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Imidazóis/farmacologia , Inflamação/genética , Lipopolissacarídeos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação , Inibidores de Proteínas Quinases/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Piridinas/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores Estrogênicos/metabolismo , Receptores Acoplados a Proteínas-G/metabolismo , Ovinos , Transdução de Sinais/efeitos dos fármacos
9.
PLoS One ; 15(8): e0237666, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32822385

RESUMO

Before fertilization, sperm bind to epithelial cells of the oviduct isthmus to form a reservoir that regulates sperm viability and capacitation. The sperm reservoir maintains optimum fertility in species, like swine, in which semen deposition and ovulation may not be well synchronized. We demonstrated previously that porcine sperm bind to two oviductal glycan motifs, a biantennary 6-sialylated N-acetyllactosamine (bi-SiaLN) oligosaccharide and 3-O-sulfated Lewis X trisaccharide (suLeX). Here, we assessed the ability of these glycans to regulate sperm Ca2+ influx, capacitation and affect sperm lifespan. After 24 h, the viability of sperm bound to immobilized bi-SiaLN and suLeX was higher (46% and 41% respectively) compared to viability of free-swimming sperm (10-12%). Ca2+ is a central regulator of sperm function so we assessed whether oviduct glycans could affect the Ca2+ influx that occurs during capacitation. Using a fluorescent intracellular Ca2+ probe, we observed that both oviduct glycans suppressed the Ca2+ increase that occurs during capacitation. Thus, specific oviduct glycans can regulate intracellular Ca2+. Because the increase in intracellular Ca2+ was suppressed by oviduct glycans, we examined whether glycans affected capacitation, as determined by protein tyrosine phosphorylation and the ability to undergo a Ca2+ ionophore-induced acrosome reaction. We found no discernable suppression of capacitation in sperm bound to oviduct glycans. We also detected no effect of oviduct glycans on sperm motility during capacitation. In summary, LeX and bi-SiaLN glycan motifs found on oviduct oligosaccharides suppress the Ca2+ influx that occurs during capacitation and extend sperm lifespan but do not affect sperm capacitation or motility.


Assuntos
Cálcio/metabolismo , Oviductos/metabolismo , Polissacarídeos/metabolismo , Espermatozoides/metabolismo , Suínos/fisiologia , Animais , Sobrevivência Celular , Feminino , Masculino , Análise do Sêmen , Capacitação Espermática , Motilidade Espermática
10.
PLoS One ; 15(8): e0237108, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32750076

RESUMO

Anatomically terminal parts of the urinary, reproductive, and digestive systems of birds all connect to the cloaca. As the feces drain through the cloaca in chickens, the cloacal bacteria were previously believed to represent those of the digestive system. To investigate similarities between the cloacal microbiota and the microbiota of the digestive and reproductive systems, microbiota inhabiting the colon, cloaca, and magnum, which is a portion of the chicken oviduct of 34-week-old, specific-pathogen-free hens were analyzed using a 16S rRNA metagenomic approach using the Ion torrent sequencer and the Qiime2 bioinformatics platform. Beta diversity via unweighted and weighted unifrac analyses revealed that the cloacal microbiota was significantly different from those in the colon and the magnum. Unweighted unifrac revealed that the cloacal microbiota was distal from the microbiota in the colon than from the microbiota in the magnum, whereas weighted unifrac revealed that the cloacal microbiota was located further away from the microbiota in the magnum than from the microbiota inhabiting the colon. Pseudomonas spp. were the most abundant in the cloaca, whereas Lactobacillus spp. and Flavobacterium spp. were the most abundant species in the colon and the magnum. The present results indicate that the cloaca contains a mixed population of bacteria, derived from the reproductive, urinary, and digestive systems, particularly in egg-laying hens. Therefore, sampling cloaca to study bacterial populations that inhabit the digestive system of chickens requires caution especially when applied to egg-laying hens. To further understand the physiological role of the microbiota in chicken cloaca, exploratory studies of the chicken's cloacal microbiota should be performed using chickens of different ages and types.


Assuntos
Galinhas/microbiologia , Cloaca/microbiologia , Colo/microbiologia , Microbioma Gastrointestinal , Oviductos/microbiologia , Animais , Feminino , Flavobacterium/genética , Flavobacterium/patogenicidade , Lactobacillus/genética , Lactobacillus/patogenicidade , Metagenoma , Pseudomonas/genética , Pseudomonas/patogenicidade
11.
C R Biol ; 343(1): 89-99, 2020 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-32720491

RESUMO

In the present study we hypothesize that aquaporin 4 (AQP4) expression in the chicken oviduct would change during a pause in egg laying that was induced by fasting. Accordingly, the aim of this investigation was to examine the AQP4 mRNA and protein expression, and immunolocalization in the chicken oviduct during the course of regression. The experiment was carried out on laying hens subjected to a pause in laying that was induced by food deprivation for 5 days. Control hens were fed ad libitum. The birds were sacrificed on day 6 of the experiment and all segments of the oviduct were isolated, including the infundibulum, magnum, isthmus, shell gland, and vagina. Subsequently, the gene and protein expressions of AQP4 in the tissues were tested by real-time PCR and Western blot, respectively. The relative mRNA expression of AQP4 was the highest in the infundibulum and vagina and the lowest, and least detectable, in the magnum. The level of AQP4 protein was the highest in the infundibulum and the lowest in the magnum. Fasting resulted in a decrease of the AQP4 mRNA expression (P<0.001) in the infundibulum, a decrease in protein abundance (P<0.01) in the shell gland, and an increase in protein level (P<0.001) in the vagina. Immunohistochemistry demonstrated tissue- and cell-dependent localization of AQP4 protein in the oviductal wall. The intensity of staining was as follows: the infundibulum > shell gland > vagina ≥ isthmus ≫ magnum. In the control hens, the immunoreactivity for AQP4 in the vagina was similar, whereas in other oviductal segments, the immunoreactivity was stronger when compared with the chickens subjected to a pause in laying. In summary, these findings suggest that the AQP4 is an essential protein involved in the regulation of water transport required to create a proper microenvironment for fertilization and egg formation in the hen oviduct.


Assuntos
Aquaporina 4/metabolismo , Galinhas/fisiologia , Privação de Alimentos , Animais , Galinhas/genética , Feminino , Humanos , Imuno-Histoquímica , Oviductos/fisiologia , Oviposição , RNA Mensageiro/genética
12.
Avian Dis ; 64(2): 149-156, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32550615

RESUMO

Infection of the oviduct by an infectious bronchitis virus (IBV) in laying hens has been associated with the false layer syndrome. Because the diagnostic procedure for the detection of cystic oviducts by postmortem examinations in IBV-positive replacement pullet flocks could involve the unnecessary sacrifice of numerous healthy pullets without reproductive tract anomalies, the development of a noninvasive and nonlethal diagnostic procedure would be desirable. The first objective of the study was to evaluate the diagnostic accuracy of a transcutaneous ultrasonography method to predict the presence of cystic oviducts compared to postmortem examinations in a commercial pullet flock positive for an IBV genotype Delmarva (DMV) variant. The second objective was to evaluate the performance of the same ultrasonography method to later detect false layers in the same flock in sexually mature hens by identifying the presence of an egg in the oviduct due to the presence of atretic oviducts undetectable by ultrasonography and the absence of cystic oviducts at that age. In replacement pullets, the sensitivity (Se) and specificity (Sp) of the ultrasonography (index test) compared to the postmortem examination (reference standard test) were 73% and 91%, respectively. The positive predictive value (PPV) and negative predictive value (NPV) were 67% and 93%. The ultrasonography technique showed a positive likelihood ratio (LR+) of 7.82 and a negative likelihood ratio (LR-) of 0.30. In sexually mature hens, the Se, Sp, PPV, and NPV of the ultrasonography compared to the laying status were 98%. The LR+ was 49.00 and the LR- was 0.02 when compared to the laying status. In conclusion, the ultrasonography could replace postmortem examinations to detect cystic oviducts in commercial flocks of replacement pullets previously infected with an IBV-DMV 1639 variant. Although the test accuracy of ultrasonography was excellent for the hens at production peak to identify laying and nonlaying hens based on the presence of an egg in the reproductive tract, its practicality was limited due to atretic oviducts being not detectable.


Assuntos
Galinhas , Infecções por Coronavirus/veterinária , Cistos/veterinária , Vírus da Bronquite Infecciosa/isolamento & purificação , Doenças das Aves Domésticas/diagnóstico , Ultrassonografia/veterinária , Animais , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/virologia , Cistos/virologia , Feminino , Oviductos/virologia , Doenças das Aves Domésticas/virologia , Ultrassonografia/estatística & dados numéricos
14.
Anim Sci J ; 91(1): e13396, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32468659

RESUMO

The objective of this study was to examine the expression profiles of follistatin (FST) and its associated molecules (MSTN, INHA, INHBB, INHBA, ACVR2A, and ACVR2B) in the oviduct of laying hens at 3 hr and 20 hr post-ovulation (p.o., n = 5; 35 weeks old), molting (n = 5; 60 weeks old), and non-laying (n = 4; 35-60 weeks old) hens and also to localize the FST by using immunohistochemistry assay. Expression of FST was significantly higher (p < .05), and MSTN was lower in the uterus of laying hens around 15-20 hr p.o. (during eggshell formation), however, their expressions in the magnum remain unchanged across different physiological stages of hens. FST was mainly expressed in the luminal and glandular epithelium of the uterine tissues, and their expression intensity was highest in laying hens during the eggshell mineralization. There was a relatively increased expression of INHA in the magnum of laying hens around 3 hr p.o. as compared to non-laying and molting hens. At the same time (3 hr p.o.), there was a significant (p < .05) decrease in the expression of the INHBB, ACVR2A, and ACV2B. These results indicate that follistatin may regulate the differentiation of uterine luminal and glandular epithelium during eggshell biomineralization.


Assuntos
Biomineralização/genética , Galinhas/genética , Galinhas/fisiologia , Casca de Ovo/embriologia , Folistatina/genética , Folistatina/metabolismo , Expressão Gênica/genética , Oviductos/metabolismo , Oviposição/genética , Oviposição/fisiologia , Ovulação/genética , Ovulação/fisiologia , Transcriptoma , Animais , Biomineralização/fisiologia , Diferenciação Celular/genética , Diferenciação Celular/fisiologia , Casca de Ovo/fisiologia , Células Epiteliais/metabolismo , Células Epiteliais/fisiologia , Feminino , Oviductos/fisiologia , Útero/citologia , Útero/metabolismo
15.
Am J Physiol Endocrinol Metab ; 318(6): E981-E994, 2020 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-32315215

RESUMO

Chlamydia trachomatis infection is a primary cause of reproductive tract diseases including infertility. Previous studies showed that this infection alters physiological activities in mouse oviducts. Whether this occurs in the uterus and cervix has never been investigated. This study characterized the physiological activities of the uterine horn and the cervix in a Chlamydia muridarum (Cmu)-infected mouse model at three infection time points of 7, 14, and 21 days postinfection (dpi). Cmu infection significantly decreased contractile force of spontaneous contraction in the cervix (7 and 14 dpi; P < 0.001 and P < 0.05, respectively), but this effect was not observed in the uterine horn. The responses of the uterine horn and cervix to oxytocin were significantly altered by Cmu infection at 7 dpi (P < 0.0001), but such responses were attenuated at 14 and 21 dpi. Cmu infection increased contractile force to prostaglandin (PGF2α) by 53-83% in the uterine horn. This corresponded with the increased messenger ribonucleic acid (mRNA) expression of Ptgfr that encodes for its receptor. However, Cmu infection did not affect contractions of the uterine horn and cervix to PGE2 and histamine. The mRNA expression of Otr and Ptger4 was inversely correlated with the mRNA expression of Il1b, Il6 in the uterine horn of Cmu-inoculated mice (P < 0.01 to P < 0.001), suggesting that the changes in the Otr and Ptger4 mRNA expression might be linked to the changes in inflammatory cytokines. Lastly, this study also showed a novel physiological finding of the differential response to PGE2 in mouse uterine horn and cervix.


Assuntos
Infecções por Chlamydia/fisiopatologia , Chlamydia muridarum , Miométrio/fisiopatologia , Infecções do Sistema Genital/fisiopatologia , Contração Uterina/fisiologia , Útero/fisiopatologia , Animais , Colo do Útero/metabolismo , Colo do Útero/fisiopatologia , Infecções por Chlamydia/genética , Infecções por Chlamydia/imunologia , Infecções por Chlamydia/metabolismo , Citocinas/genética , Dinoprosta/farmacologia , Dinoprostona/farmacologia , Feminino , Regulação da Expressão Gênica , Histamina/farmacologia , Agonistas dos Receptores Histamínicos/farmacologia , Interleucina-1beta/genética , Interleucina-6/genética , Camundongos , Contração Muscular/efeitos dos fármacos , Contração Muscular/fisiologia , Músculo Liso/efeitos dos fármacos , Músculo Liso/fisiopatologia , Miométrio/efeitos dos fármacos , Miométrio/metabolismo , Oviductos/patologia , Ocitócicos/farmacologia , RNA Mensageiro/metabolismo , Receptores de Ocitocina/genética , Receptores de Prostaglandina/genética , Receptores de Prostaglandina E Subtipo EP4/genética , Infecções do Sistema Genital/genética , Infecções do Sistema Genital/imunologia , Infecções do Sistema Genital/metabolismo , Contração Uterina/efeitos dos fármacos , Útero/metabolismo
16.
Int J Mol Sci ; 21(5)2020 Feb 29.
Artigo em Inglês | MEDLINE | ID: mdl-32121434

RESUMO

Knowledge of how the biochemical composition of the bovine oviduct is altered due to the oviduct anatomy or the presence of an embryo is lacking. Thus, the aim of this study was to assess the effect of (І) oviduct anatomy and (ІІ) embryo presence on oviductal fluid (OF) protein, amino acid, and carbohydrate composition. Cross-bred beef heifers (n = 19) were synchronized and those in standing estrus were randomly allocated to a cyclic (non-bred) or pregnant (artificially inseminated) group. All heifers were slaughtered on Day 3 after estrus. The oviducts ipsilateral to the corpus luteum from each animal were isolated, straightened and cut, separating ampulla and isthmus. Each portion was flushed with 500 µl of PBS enabling recovery of the oocyte/embryo. Recovered unfertilized oocytes (cyclic group) and embryos (8-cell embryos; pregnant group) were located in the isthmus of the oviduct. Samples of flushing medium from the isthmus and ampulla were used for proteomic (n = 2 per group), amino acid (n = 5), and carbohydrate (n = 5) analysis. For proteomic analysis, total protein from cyclic and pregnant samples were labelled with different cyanine fluorescent probes and separated according to the isoelectric point using immobilized pH gradient strips (pH 3-10, 17 cm, Protean® IEF cell system, Bio Rad). Second dimension was performed in a polyacrylamide gel (12%) in the presence of SDS using a Protean II XL system (Bio Rad). Images were obtained with a Typhoon 9410 scanner and analyzed with Progenesis SameSpots software v 4.0. Amino acid content in the OF was determined by high performance liquid chromatography (HPLC). Glucose, lactate, and pyruvate were quantified using microfluorometric enzyme-linked assays. For the proteomic assessment, the results of the image analysis were compared by ANOVA. For both amino acid and carbohydrate analyses, statistical analysis was carried out by 2-way ANOVA with the Holm-Sidak nonparametric post hoc analysis. On Day 3 post-estrus, OF composition varied based on (І) anatomical region, where isthmic metabolites were present in lower (i.e., lactate, glycine, and alanine) or higher (i.e., arginine) concentrations compared to the ampulla; and (ІІ) embryo presence, which was correlated with greater, arginine, phosphoglycerate kinase 1, serum albumin, α-1-antiproteinase and IGL@ protein concentrations. In conclusion, data indicate that the composition of bovine OF is anatomically dynamic and influenced by the presence of an early embryo.


Assuntos
Aminoácidos/genética , Metabolismo dos Carboidratos/genética , Proteínas/genética , Proteômica , Aminoácidos/metabolismo , Animais , Bovinos , Embrião de Mamíferos , Tubas Uterinas/metabolismo , Feminino , Oócitos/metabolismo , Oviductos/metabolismo , Gravidez , Proteínas/metabolismo
17.
Poult Sci ; 99(3): 1705-1716, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32115038

RESUMO

Vanadium (V) is an ultratrace metal with the insulin-tropic properties and is often researched as the diabetes drug. However, in animals, V has been reported to have toxic effects on the development, immunity, oxidation-reduction equilibrium, gastrointestinal function, and so forth. Especially in poultry, supplementation of more than 10 mg of V/kg in the layer diets has been shown to adversely affect the egg production and egg quality. In this study, we supplemented 0 mg of V/kg, 5 mg of V/kg, and 10 mg of V/kg in the layer diets for 35 D and examined the quantitative proteomics of albumen for finding the possible target signaling pathway and mechanism of V action and made the preliminary verification. In contrast to the control group, V resulted in a significant drop in the albumen height, and in oviduct ampulla, the activity of total antioxidant capacity and glutathione peroxidase significantly decreased (P = 0.01, P = 0.02), the content of malonic dialdehyde significantly increased (P = 0.01), and the apoptosis rate significantly increased in the 5-mg V/kg and 10-mg V/kg treatment groups (P < 0.01). V affected 36 differentially accumulated proteins in albumen, with 23 proteins upregulated and 13 proteins downregulated. The expressions of innate protein albumen lysozyme (Q6LEL2), vitellogenin-2 (P02845), and the F1NWD0 protein in albumen belonged to the P53 family were significantly reduced, in contrast to the control (P < 0.05), and the expression of riboflavin-binding protein (P02752) was significantly improved (P < 0.05). The Hippo signaling pathway-fly, which is suitable for the key protein P53 as the most significantly affected network, might be important for discriminating V.


Assuntos
Ração Animal/análise , Clara de Ovo/análise , Proteoma , Vanádio/efeitos adversos , Animais , Galinhas , Dieta/veterinária , Proteínas do Ovo/química , Feminino , Oviductos , Transdução de Sinais
18.
J Toxicol Sci ; 45(3): 131-136, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32147636

RESUMO

Reproductive disorders in birds are the most characteristic effects of DDT contamination of wildlife. Experimental exposure of avian eggs to the estrogenic substance o,p'-DDT causes abnormal development of the reproductive tract (shortening of the left oviduct and aberrant development of the right oviduct) and eggshell thinning in mature birds, but it is still not known how eggshell thinning occurs in the abnormal oviduct. To fill this information gap, we examined the histology of the uterine part of the oviduct in Japanese quail treated in ovo with o,p'-DDT or a synthetic estrogen, diethylstilbestrol (DES), and we performed immunohistochemical staining for the calcium-binding proteins CALB1, SPP1, and TRPV6. Both o,p'-DDT-treated and DES-treated quail had few, and scattered, gland cells in the left uterus, unlike vehicle controls, in which gland cells tightly occupied the lamina propria. The aberrantly developed right uterus retained all the components of the normal left uterus, but in immature form. Immunostaining for CALB1, SPP1, and TRPV6 was greatly reduced by both o,p'-DDT and DES; SPP1 and TRPV6 immunostaining patterns, in particular, differed distinctly from those in the controls. These findings suggest that CALB1, SPP1, and TRPV6 are molecular factors, decreased production of which is responsible for eggshell thinning. Our findings also could contribute to understanding of the eggshell formation mechanism in birds.


Assuntos
Proteínas de Ligação ao Cálcio/metabolismo , DDT/toxicidade , Casca de Ovo/efeitos dos fármacos , Oviductos/efeitos dos fármacos , Oviductos/metabolismo , Animais , Coturnix , Dietilestilbestrol/toxicidade , Casca de Ovo/patologia , Feminino , Oviductos/patologia
19.
Int J Mol Sci ; 21(4)2020 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-32075098

RESUMO

Oviductal extracellular vesicles (oEVs) are emerging as key players in the gamete/embryo-oviduct interactions that contribute to successful pregnancy. Various positive effects of oEVs on gametes and early embryos have been found in vitro. To determine whether these effects are associated with changes of embryonic gene expression, the transcriptomes of embryos supplemented with bovine fresh (FeEVs) or frozen (FoEVs) oEVs during in vitro culture compared to controls without oEVs were analyzed by low-input RNA sequencing. Analysis of RNA-seq data revealed 221 differentially expressed genes (DEGs) between FoEV treatment and control, 67 DEGs for FeEV and FoEV treatments, and minor differences between FeEV treatment and control (28 DEGs). An integrative analysis of mRNAs and miRNAs contained in oEVs obtained in a previous study with embryonic mRNA alterations pointed to direct effects of oEV cargo on embryos (1) by increasing the concentration of delivered transcripts; (2) by translating delivered mRNAs to proteins that regulate embryonic gene expression; and (3) by oEV-derived miRNAs which downregulate embryonic mRNAs or modify gene expression in other ways. Our study provided the first high-throughput analysis of the embryonic transcriptome regulated by oEVs, increasing our knowledge on the impact of oEVs on the embryo and revealing the oEV RNA components that potentially regulate embryonic development.


Assuntos
Embrião de Mamíferos/metabolismo , Vesículas Extracelulares/metabolismo , RNA/metabolismo , Transcriptoma , Animais , Bovinos , Análise por Conglomerados , Embrião de Mamíferos/citologia , Desenvolvimento Embrionário/genética , Feminino , Congelamento , Regulação da Expressão Gênica no Desenvolvimento , MicroRNAs/metabolismo , Oviductos/citologia , Oviductos/metabolismo , RNA/química , RNA Mensageiro/metabolismo , Análise de Sequência de RNA
20.
Int J Mol Sci ; 21(3)2020 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-32050564

RESUMO

Embryo-maternal crosstalk is an important event that involves many biological processes, which must occur perfectly for pregnancy success. This complex communication starts from the zygote stage within the oviduct and continues in the uterus up to the end of pregnancy. Small extracellular vesicles (EVs) are part of this communication and carry bioactive molecules such as proteins, lipids, mRNA, and miRNA. Small EVs are present in the oviductal and uterine fluid and have important functions during fertilization and early embryonic development. Embryonic cells are able to uptake oviductal and endometrium-derived small EVs. Conversely, embryo-derived EVs might modulate oviductal and uterine function. In this review, our aim is to demonstrate the role of extracellular vesicles modulating embryo-maternal interactions during early pregnancy.


Assuntos
Blastocisto/metabolismo , Comunicação Celular , Endométrio/metabolismo , Vesículas Extracelulares/metabolismo , Animais , Feminino , Humanos , Oviductos/metabolismo , Zigoto/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...